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Many infectious diseases can fade away after initial outbreaks. Bubonic plague, cholera, and influenza are examples from recent human history. The same phenomenon occurs for wildlife diseases as well. How does this happen? One popular explanation is that the pathogen evolves to become less deadly, so that it doesn’t completely wipe out its hosts, ensuring pathogen survival. While this scenario does sometimes play out, we know that there are other reasons why the severity of diseases can change over time.

Cori Richards and Jamie Voyles

For amphibians, we’ve known about a highly lethal disease called “chytridiomycosis” since the 1990s. This disease was especially devastating in Central America, where it may have wiped out entire species. In this study, we made the exciting discovery that some amphibian species – frogs that were thought to be extinct – are persisting, and even recovering, after lethal disease outbreaks. We wanted to understand how it was happening. Was it a change in the pathogen, the frogs, or both?

To answer these questions, we did two things. To begin with, we surveyed frogs in Panama before and after the disease outbreak. In addition, we collected samples of the pathogen at multiple time points: during initial outbreaks and ~10 years later. We found that nearly a decade after the outbreak, the pathogen was just as deadly. However, the frogs are surviving and have better defenses against it. Panama’s frogs are fighting back! Understanding how amphibian communities are recovering after this disease outbreak is important multiple reasons. First, resolving how this works will help us develop more informed conservation strategies to protect amphibians from disease-induced extinctions. Second, clarifying how disease outbreaks subside will help us predict, and respond to, other emerging pathogens in plants, wildlife, and in humans. These goals are increasingly important in a time when rapid globalization has increased the introduction of pathogens to naïve host populations.

Atelopus varius is one species that appears to have evolved antifungal skin secretions

A recent rule put in place in 2016, restricting the international import of 201 salamander species into the United States, aimed to prevent the newly discovered deadly salamander fungal disease, Batrachochytrium salamandrivorans (Bsal), from entering the country. In a new study published Oct. 13 in Scientific Reports, Smithsonian Conservation Biology Institute scientists reveal that the moratorium seemingly has a chance to do its job effectively.

Researchers swabbing an emperor newt at the Smithsonian’s National Zoo. Emperor newts belong to a genus of newts from Asia that are currently subjected to the U.S. Fish and Wildlife Service’s moratorium on salamander imports because of the risk that they may carry the deadly salamander fungal disease, Batrachochytrium salamandrivorans (Bsal).

“When the moratorium went into effect, we did not know if Bsal was already in the United States in pet salamanders and whether we were closing the barn door after the horse had already escaped,” said Brian Gratwicke, SCBI amphibian conservation biologist and paper senior author. “Our study did not find the pathogen in pet salamander populations in the United States, which is good news for native salamanders, especially in the Appalachian region—a salamander biodiversity hotspot. It also means that we must continue to be vigilant and prevent the disease from entering the country.”

The study marks the first general survey for Bsal in pet salamanders in the United States. The researchers worked with the Amphibian Survival Alliance to mail out sampling kits to salamander pet owners. In return, the team received skin swab samples from 639 salamanders belonging to 65 species, many of which are potential carriers of Bsal. None of the samples came back with evidence of Bsal, according to tests conducted in SCBI’s Center for Conservation Genomics.

“Working with the pet-hobbyist community on this project gave us a chance to alert this key group to a potential problem and was critical in determining whether Bsal has been imported into the United States,” said Blake Klocke, George Mason University’s Department of Environmental Science and Policy doctoral student, researching with SCBI and lead author on the study. “We hope that they will continue to be watchdogs for signs of Bsal and will implement testing and biosecurity protocols into their regular routine to prevent the possible spread of disease in the future.”

Bsal was discovered after populations of fire salamanders in the Netherlands experienced catastrophic declines from the disease, which was likely introduced from Asia, the source of most international exports of salamander species for the pet trade. Bsal is similar to a frog-killing fungus called Batrachochytrium dendrobatidis (Bd), which has been a major driver of global amphibian declines and extinctions. Bsal has been detected in the wild in the Netherlands, Belgium, Germany and Vietnam, as well as in in captive individuals in the United Kingdom and Germany.

The Lacey Act, which includes the 201 species of salamanders the U.S. Fish and Wildlife Service list as “injurious wildlife” (those most susceptible to Bsal or likely to spread Bsal) limits both the import of these animals from other countries and their transfer over state lines. According to the paper, the Lacey Act decision reduced the number of salamanders imported to the United States from 2015 to 2016 by 98.4 percent.

The United States is home to 190 native species of salamanders. The Scientific Reports study complements SCBI’s ongoing tests of salamanders in the wild, which have also come back negative for Bsal. SCBI will continue to screen for the disease in the wild and work with collaborators on developing methods to manage the spread of Bsal should it be introduced into the wild.

“Salamanders play a key role in maintaining the health of our forests and may even help regulate climate,” said Carly Muletz-Wolz, SCBI research scientist and paper co-author. “If Bsal were to hitch a ride to the eastern United States specifically, where salamanders are particularly abundant, it could spread quickly and result in catastrophic changes to the ecosystems. It is imperative that we do all we can to prevent the introduction of Bsal into the country and that we continue to monitor our wild populations so we can take swift action if needed.”

The paper’s additional authors are Matthew Becker and Robert Fleischer, SCBI; James Lewis, Rainforest Trust; and Larry Rockwood and A. Alonso Aguirre, George Mason University.

What’s Working in Conservation

The global conservation movement has reached a turning point. We have documented the fast pace of habitat loss, the growing number of endangered and extinct species, and the increasing speed of global climate change. Yet while the seriousness of these threats cannot be denied, there are a growing number of examples of improvements in the health of species and ecosystems, along with benefits to human well-being, thanks to our conservation actions. Earth Optimism is a global initiative that celebrates a change in focus from problem to solution, from a sense of loss to one of hope, in the dialogue about conservation and sustainability.

As human diseases become alarmingly antibiotic resistant, identification of new pharmaceuticals is critical. The cane toad and other members of the Bufonidae family produce substances widely used in traditional folk medicine, but endangered family members, like Panama’s golden frog, Atelopus zeteki, may disappear before revealing their secrets. Smithsonian scientists and colleagues catalog the known chemicals produced by this amphibian family in the Journal of Ethnopharmacology highlighting this largely-unexplored potential for discovery.

“We’re slowly learning to breed members of this amphibian family decimated by the chytrid fungal disease,” said Roberto Ibañez, Panamanian staff scientist at the Smithsonian Tropical Research Institute (STRI) and in-country director of the Panama Amphibian Conservation and Rescue (PARC) project. “That’s buying us time to understand what kind of chemicals they produce, but it’s likely that animals in their natural habitats produce an even wider range of compounds.”

15 of 47 frog and toad species used in traditional medicine belong to the family Bufonidae. For millennia, secretions from their skin and from glands near their ears called parotid glands, as well as from their bones and muscle tissues have been used as remedies for infections, bites, cancer, heart disorders, hemorrhages, allergies, inflammation, pain and even AIDS.

Toxins of two common Asian toad species, Bufo gargarizans and Duttaphrynus melanostictus, produce the anticancer remedies known as Chan Su and Senso in China and Japan, respectively. Another preparation used to treat cancer and hepatitis, Huachansu or Cinobufacini, is regulated by the Chinese State Food and Drug Administration. In Brazil, the inner organs of the toad, Rhinella schneideri, are applied to horses to treat the parasite Habronema muscae. In Spain, extract from the toad Bufo bufo is used to treat hoof rot in livestock. In China, North and South Korea, ranchers use the meat of Bufo gargarizans to treat rinderpest.

Only a small proportion of the more than 580 species in the Bufonidae family have been screened by scientists. “In Panama, not only do we have access to an amazing diversity of amphibian species,” said Marcelino Gutiérrez, investigator at the Center for Biodiversity and Drug Discovery at Panama’s state research institute, Instituto de Investigaciones Cientificas y Servicios de Alta Tecnologia (INDICASAT), “we’re developing new mass spectrometry and nuclear magnetic resonance spectroscopy techniques to make it easier and cheaper to elucidate the chemical structures of the alkaloids, steroids, peptides and proteins produced by these animals. We work closely with herpetologists so as not to further threaten populations of these species in the wild.” Their efforts to catalog chemicals produced by the Bufonidae included researchers from the University of Panama, Vanderbilt University, in Tennessee, U.S.A. and Acharya Nagarjuna University in Guntur, India.

Most of the chemicals produced by frogs and toads protect them against predators. Atelopus varius contains tetrodototoxin. Chiriquitoxin is found in Atelopus limosus, one of the first species that researches succeeded in breeding in captivity as well as in Atelopus glyphus and Atelopus chiriquiensis. An atelopidtoxin (zetekitoxin) from the Panamanian golden frog, Atelopus zeteki, appears to consist of two toxins. Toxins from a single frog skin can kill 130-1000 mice.

The golden frog, A. zeteki, Panama’s national frog, is the only species of the genus Atelopus that secretes zetekitoxins. Threatened by the chytrid fungal disease that infects the skin and causes heart attacks, with collection for the exotic pet trade and by habitat destruction, if golden frogs were to disappear, they would take this potentially valuable chemical with them.

More than 30 percent of amphibians in the world are in decline. Racing to stay ahead of the wave of disease spreading across Central America, Panama is leading the way in conservation efforts. The Smithsonian’s Panama Amphibian Rescue and Conservation project (PARC) identified several Atelopus species in danger of extinction, and are learning how to create the conditions needed to breed them in captivity. Not only do animal caretakers at their facilities in Gamboa and El Valle, Panama experiment to discover what the frogs eat, they also recreate the proper environment the entire frog life-cycle: egg laying, egg hatching and tadpole survival, to successfully breed Atelopus. Each species has unique requirements, making it an expensive challenge to create this Noah’s ark for amphibians.

The chemical building blocks amphibians use to create toxic compounds come from sources including their diet, skin glands or symbiotic microorganisms. Toads in the genus Melanophryniscus sequester lipophilic alkaloids from their complex diet consisting of mites and ants. Researchers found that toxins found in a wild-caught species of Atelopus could not be isolated from frogs raised in captivity: another reason to conserve frog habitat and to begin to explore the possibility of releasing frogs bred in captivity back into the wild.

Learn more about amphibians by visiting the PARC blog and the Panama’s Fabulous Frogs exhibit at the Smithsonian’s Culebra Point Nature Center in Panama.

The Smithsonian Tropical Research Institute, headquartered in Panama City, Panama, is a part of the Smithsonian Institution. The Institute furthers the understanding of tropical nature and its importance to human welfare, trains students to conduct research in the tropics and promotes conservation by increasing public awareness of the beauty and importance of tropical ecosystems. Website. Promo video.

For five years, I have worked as the staff herpetologist at Las Casas de la Selva, a sustainable forestry project in Puerto Rico. I’ve also had the privilege of spending five months in Panama; some of that time was spent working with PARC and I have noticed many similarities between Puerto Rico and Panama. Specifically, I have noticed how each country has a frog as a national and cultural symbol.

In Panama, everyone knows of the Golden Frog (Atelopus zeteki), and when I was studying the folklore of Panama, I heard old stories about how the frog was good luck, and that people used to believe that it turned to gold when it died. Nowadays, the Golden Frog is a symbol of Panama itself, and what it means to be Panamanian. Images of the Golden Frog adorn everything from lottery tickets to t-shirts to coffee mugs. Some of my friends in Panama have even gone so far as to get Golden Frog tattoos. And each year, thousands of people celebrate Golden Frog Day.

As an island, Puerto Rico has very few species of frogs, and 16 of the 18 native species belong to the genus Eleutherodactylus. This genus is referred to collectively as “Coquis”, although only two species make the distinctive “Ko-Kee” mating call that makes nighttime in Puerto Rico such a noisy affair. Of the 16 species of Coquis, 13 are listed by the IUCN as either Vulnerable, Endangered, or Critically Endangered. As with the Golden Frog in Panama, Coquis are a symbol of everything Puerto Rican. Mainland-born Puerto Ricans who return to the island respond to challenges about their Puerto Rican “authenticity” used to respond, “I’m as Puerto Rican as the Coqui”. Images of the Coqui show up on artistic murals, tourist kitch, and tattoos; there is even a “Coqui” brand of coffee and a “Coqui” car dealership!

But we are missing something in Puerto Rico- we have no equivalent to the “Dia de Rana Dorada”. After my time at PARC, including my opportunity in 2012 to help Angie Estrada, Jorge Guerrel, Ligo Diaz, and the rest of the staff plan and execute educational activities at the Summit Zoo, I decided to take the spirit of “Dia de Rana Dorada” back to Puerto Rico with me. The idea has been well-received, and the first “Dia del Coqui” will be a weekend-long festival from September 26th-28th, 2014. It will be held at the Jardin Botanical y Cultural William Miranda Marin in the centrally-located city of Caguas. Already, artisans, scientists, musicians, university students, historians, and public-works officials are coming together to make this event a success.

The intention of Dia del Coqui is to be a cultural celebration, but also an important learning tool to help the people of Puerto Rico know that the frogs that they have always shared the island with are in need of conservation. We hope that Dia del Coqui becomes a cultural mainstay in Puerto Rico, akin to Dia de Rana Dorada in Panama.

There are two types of golden frogs in Panama. This one, Atelopus varius is the variable harlequin frog, which ranges in appearance from from this browner form to a bright yellow color at some sites. Atelopus zeteki is the distinctive day-glow yellow Panamanian golden frog. Photo: Jamie Voyles, Project Atelopus

Jamie Voyles

One small frog can offer a great deal of hope. This frog, an adult male Atelopus varius, belongs to a genus that is critically endangered – not a single species, but the entire genus – and it is, therefore, one of the most rare creatures on earth. The Panamanian golden frog Atelopus zeteki carries the additional distinction of being Panama’s national animal and it is a symbol of good luck for the Panamanian people – so much so, that in the past golden frogs graced the face of lottery tickets. So, the loss of Atelopus, due to the lethal disease chytridiomycosis, has been nothing short of a tragedy for Panamanians, as well as for the larger global community.

About a decade ago, together with my colleague, Cori Richards (now Dr. Cori Richards-Zawacki), and I watched these golden jewels vanish from the streams of Panama as the disease chytridiomycosis (“chytrid”) spread across the country. Cori and I were still graduate students; we had a youthful (albeit slightly naïve) enthusiasm for confronting the ominous conservation disaster. Cori focused on golden frogs for her PhD and sampled thousands of frogs before they succumbed to disease. I was interested in understanding which species would be affected by the disease, not knowing that chytrid would, in a few short years, cause a wholesale wipe-out of entire amphibian communities. When frogs started to disappear, our research projects ground to a halt. After all, no frogs means no frog research. So our advisers, perhaps wisely, advised us to move on. As we shifted our research projects to other locales, the golden frogs reached the brink of extinction; sightings of these now-rare creatures dwindled until they were mere rumors.

Fast-forward ten years. Cori and I had both advanced in our academic careers, but we were still haunted by the loss of Panamanian amphibians. When Cori visited the University of California, Berkeley, where I was finishing up my post-doctoral work, we spent an afternoon sitting on the green campus lawn and reflecting on our work of a decade earlier; despite the projections, we had not realized the full scope of what had been coming – especially for those beautiful Panamanian frogs. In those days, we remembered, not very many people outside of a small group of researchers had even heard the word “chytrid”, much less tried to pronounce it. Few were paying attention to the global decline of amphibian populations; even fewer were aware of Panama’s devastating loss of its national mascot and lucky charm. So, naturally, we hatched a plan to return. We needed to see for ourselves what remained of the Panama’s golden frogs.

We set about gathering money from conservation grants, one small award at a time. The news coming from field reports was grim, but we remained determined. We pooled our small pots of funds (including support from the Smithsonian and Project Golden Frog) and galvanized small research team (including Edgardo Griffiths, Heidi Ross and Matt Robak). Soon enough, we trudged the misty mountains of Panama, machetes in hand and hopes held high despite the overwhelming odds. We visited numerous sites where Atelopus varius and Atelopus zeteki were historically found, including all of Cori’s old golden frog sites. We followed rumors, tips and hints. After several months of surveys, after hours of climbing trails with heavy packs and muddy boots, we repeatedly stumbled out of the rainforest disappointed, bug-bitten and empty handed. It would have been easy to admit that we weren’t chasing frogs anymore – now we were chasing ghosts.

Until, after months of searching, we finally found our glimmer of hope. On November 8, 2012 we found a healthy adult male Atelopus perched on a mossy boulder, unconcerned that a cross-continent scramble had been underway for months, just to find him. We sat in the rain, watching him and snapping pictures. We collected non-invasive samples for diagnostic and genetic testing and then, somewhat reluctantly, we said good-bye and wished him well. We were overjoyed…. and here’s why: One small frog in the wild suggests that there are at least some surviving populations out there. And if there is even one small population holding on, there is hope – not just for that population, or even for the species, but for the whole genus. Having evidence to support that hope, in the form of that single, small and beautiful frog, is something even better than holding a winning lottery ticket with his picture on it.

With the support from USAID and the Smithsonian Tropical Research Institute, Amphibian Ark held the Amphibian Conservation Training for Latin America’s workshop in Panama last April. Twenty seven participants from Chile, Colombia, Guatemala, Ecuador, Mexico and Panama shared experiences and information about design, implementation and husbandry of ex-situ amphibian conservation programs. The main goal was to guide amphibian conservation programs in Latin America to the next level by successfully caring for and breeding endangered species in captivity, population management and developing an exit plan with possible reintroduction methods.

Participants received intense theoretical and practical training from some of the best amphibian specialists in the region. Amphibian crisis, species status, medical issues and veterinary care, food culture, daily husbandry and biosecurity standards were some of the subject discussed, complemented with group projects and nocturnal field trips around Parque Nacional Soberania.

We thank Ron Gagliardo, Amphibian Ark Trainning Officer; Luis Carrillo, Diego Almeida Reinoso, Brian Kubicki, Brad Wilson VMD, Eric Baitchman VMD, Edgardo Griffith, Roberto Ibanez, Jorge Guerrel and Angie Estrada for donating their time and sharing all their knowledge and experience. And to all the participants for their interest, enthusiasm and work on amphibian conservation.

The limosa harlequin frog (Atelopus limosus), an endangered species native to Panama, now has a new lease on life. The Panama Amphibian Rescue and Conservation Project is successfully breeding the chevron-patterned form of the species in captivity for the first time. The rescue project is raising nine healthy frogs from one mating pair and hundreds of tadpoles from another pair.

“These frogs represent the last hope for their species,” said Brian Gratwicke, international coordinator for the project and a research biologist at the Smithsonian Conservation Biology Institute, one of six project partners. “This new generation is hugely inspiring to us as we work to conserve and care for this species and others.”

Nearly one-third of the world’s amphibian species are at risk of extinction. The rescue project aims to save priority species of frogs in Panama, one of the world’s last strongholds for amphibian biodiversity. While the global amphibian crisis is the result of habitat loss, climate change and pollution, a fungal disease, chytridiomycosis, is likely responsible for as many as 94 of 120 frog species disappearing since 1980.

Between its facilities at the Smithsonian Tropical Research Institute in Gamboa, Panama, and the El Valle Amphibian Conservation Center in El Valle, Panama, the rescue project currently cares for 55 adult limosa harlequin frogs of the chevron-patterned form and 10 of the plain-color form. The project has had limited success breeding the plain-color form of this species, and has successfully bred other challenging endangered species, including crowned treefrogs (Anotheca spinosa), horned marsupial frogs (Gastrotheca cornuta) and toad mountain harlequin frogs (A. certus).

Each species requires its own unique husbandry to thrive and breed. The project’s animal care team and scientists learn husbandry techniques as they work with a limited number of individuals. Jorge Guerrel, conservation biologist at the Smithsonian Tropical Research Institute, arranged rocks in the breeding tank to create the submerged caves that appear to be the preferred egg deposition sites for limosa harlequin frogs. Like other Atelopus species, tadpoles require highly oxygenated, gently flowing water between 22 and 24 degrees Celsius. The tadpoles’ natural food is algal film growing on submerged rocks, which Guerrel and his colleagues re-created by painting petri dishes with a solution of powdered spirulina algae, then allowing it to dry.

The mission of the Panama Amphibian Rescue and Conservation Project is to rescue amphibian species that are in extreme danger of extinction throughout Panama. The project’s efforts and expertise are focused on establishing assurance colonies and developing methodologies to reduce the impact of the amphibian chytrid fungus so that one day captive amphibians may be reintroduced to the wild. Current project partners include Cheyenne Mountain Zoo, Houston Zoo, Smithsonian’s National Zoological Park, Smithsonian Tropical Research Institute and Zoo New England.

The casque headed tree frog (Hemiphractidae: Hemiphractus fasciatus), is one of 11 species of highest conservation concern now being bred in captivity in Panama. Females carry eggs on their backs where the young complete development hatching out as miniature frogs. DNA barcoding data suggest that populations of H. fasciatus may comprise more than one taxonomic group.

What if Noah got it wrong? What if he paired a male and a female animal thinking they were the same species, and then discovered they were not the same and could not produce offspring? As researchers from the Panama Amphibian Rescue and Conservation Project race to save frogs from a devastating disease by breeding them in captivity, a genetic test averts mating mix-ups.

At the El Valle Amphibian Conservation Center, project scientists breed 11 different species of highland frogs threatened by the chytrid fungus Batrachochytrium dendrobatidis, which has already decimated amphibian populations worldwide. They hope that someday they will be able to re-release frogs into Panama’s highland streams.

Different frog species may look very similar.

“If we accidentally choose frogs to breed that are not the same species, we may be unsuccessful or unknowingly create hybrid animals that are maladapted to their parents’ native environment,” said Andrew J. Crawford, research associate at the Smithsonian Tropical Research Institute in Panama and professor at Colombia’s Universidad de los Andes. Crawford and his colleagues make use of a genetic technique called DNA barcoding to tell amphibian species apart. By comparing gene sequences in a frog’s skin cells sampled with a cotton swab, they discover how closely the frogs are related.

The Smithsonian Tropical Research Institute, headquartered in Panama City, Panama, is a unit of the Smithsonian Institution. The Institute furthers the understanding of tropical nature and its importance to human welfare, trains students to conduct research in the tropics and promotes conservation by increasing public awareness of the beauty and importance of tropical ecosystems.

On Dec. 30, the Washington Post ran a front-page story about the rescue project.

The year ended on a high note for the Panama Amphibian Rescue and Conservation Project. William Booth, a science writer for the Washington Post, joined rescue project researchers on a field expedition and his story about the rescue project came out on the front page of the Post on Dec. 30. The story inspired one reader, Tim Torkildson, to share a lovely poem about frogs and the disease that is wiping them out.

If you saw the story and are interested in making a donation to the rescue project, please follow this link to the National Zoo’s website.

THE FROG
by Tim Torkildson

The frog is an amphibian
Who thrives most ev’rywhere,
From the dry Namibian
To just off ol’ Times Square.
The ones who have a bumpy skin,
With warts and pits and nodes,
Are the closest Phylum kin;
We simply call them toads.
The bullfrogs in the early spring
give ponds reverberation
With their raucous verbal fling,
Attempting procreation.
The have a courtship ritual
that’s called, I think, amplexus,
Which gives them fits conniptual
Between the two odd sexes.
A little boy will manage to
Corral a tadpole, yes,
And give it quite a slimy view
Right down his sister’s dress!
And did you know the urine from
a pregnant lady will
cause some frogs to lay a scum
of eggs, with no male thrill?
And so they’re useful critters,
As the French will tell you so;
Their legs taste good in fritters,
Are mistaken for turbot.
And what of cane toads, mind you,
Where, if you lick the skin,
The psychedelics blind you
To sorrow, grief and sin?
But frogs, those little gargoyles,
Which are funny in cartoons,
Are engaged in lethal broils
That leaves their lives in ruins.
A fungus known as “Bd” kills
The frogs down in Belize,
Then jumps the valleys and the hills
So others it may seize.
The Costa Rica Golden Toad
Is now extinct, alack.
More are headed down that road,
Since habitat is slack.
Toxins give some frogs three legs,
Which doesn’t help them jump.
Instead they are like clumsy kegs
Who in the water flump.
Scientists preserve some frogs
In habitats in labs.
Dressed in their starched, stiff white togs
They keep meticulous tabs.
To save the frogs, oh please donate
A dollar or a yuan,
So the polliwog birth rate
Will someday be a shoo-in!